Laboratory of Cellular and Molecular Immunology, Gavin Herbert Eye Institute, University of California Irvine, School of Medicine, Irvine, California, USA.
Department of Vaccines and Immunotherapies, TechImmune, LLC, University Lab Partners, Irvine, California, USA.
J Virol. 2024 May 14;98(5):e0159623. doi: 10.1128/jvi.01596-23. Epub 2024 Apr 8.
Following acute herpes simplex virus type 2 (HSV-2) infection, the virus undergoes an asymptomatic latent infection of sensory neurons of dorsal root ganglia (DRG). Chemical and physical stress cause intermittent virus reactivation from latently infected DRG and recurrent virus shedding in the genital mucosal epithelium causing genital herpes in symptomatic patients. While T cells appear to play a role in controlling virus reactivation from DRG and reducing the severity of recurrent genital herpes, the mechanisms for recruiting these T cells into DRG and the vaginal mucosa (VM) remain to be fully elucidated. The present study investigates the effect of CXCL9, CXCL10, and CXCL11 T-cell-attracting chemokines on the frequency and function of DRG- and VM-resident CD4 and CD8 T cells and its effect on the frequency and severity of recurrent genital herpes in the recurrent herpes guinea pig model. HSV-2 latent-infected guinea pigs were immunized intramuscularly with the HSV-2 ribonucleotide reductase 2 (RR2) protein () and subsequently treated intravaginally with the neurotropic adeno-associated virus type 8 expressing CXCL9, CXCL10, or CXCL11 chemokines to recruit CD4 and CD8 T cells into the infected DRG and VM (). Compared to the RR2 therapeutic vaccine alone, the RR2/CXCL11 prime/pull therapeutic vaccine significantly increased the frequencies of functional tissue-resident and effector memory CD4 and CD8 T cells in both DRG and VM tissues. This was associated with less virus in the healed genital mucosal epithelium and reduced frequency and severity of recurrent genital herpes. These findings confirm the role of local DRG- and VM-resident CD4 and CD8 T cells in reducing virus shedding at the vaginal site of infection and the severity of recurrent genital herpes and propose the novel prime-pull vaccine strategy to protect against recurrent genital herpes.IMPORTANCEThe present study investigates the novel prime/pull therapeutic vaccine strategy to protect against recurrent genital herpes using the latently infected guinea pig model. In this study, we used the strategy that involves immunization of herpes simplex virus type 2-infected guinea pigs using a recombinantly expressed herpes tegument protein-ribonucleotide reductase 2 (RR2; prime), followed by intravaginal treatment with the neurotropic adeno-associated virus type 8 expressing CXCL9, CXCL10, or CXCL11 T-cell-attracting chemokines to recruit T cells into the infected dorsal root ganglia (DRG) and vaginal mucosa (VM) (pull). We show that the RR2/CXCL11 prime-pull therapeutic vaccine strategy elicited a significant reduction in virus shedding in the vaginal mucosa and decreased the severity and frequency of recurrent genital herpes. This protection was associated with increased frequencies of functional tissue-resident (T cells) and effector (T cells) memory CD4 and CD8 T cells infiltrating latently infected DRG tissues and the healed regions of the vaginal mucosa. These findings shed light on the role of tissue-resident and effector memory CD4 and CD8 T cells in DRG tissues and the VM in protection against recurrent genital herpes and propose the prime-pull therapeutic vaccine strategy in combating genital herpes.
在急性单纯疱疹病毒 2 型（HSV-2）感染后，病毒会在背根神经节（DRG）的感觉神经元中经历无症状潜伏感染。化学和物理应激会导致潜伏感染的 DRG 间歇性病毒再激活，并导致生殖器黏膜上皮中病毒再次脱落，从而导致症状性患者发生生殖器疱疹。虽然 T 细胞似乎在控制 DRG 中的病毒再激活和减轻复发性生殖器疱疹的严重程度方面发挥作用，但将这些 T 细胞招募到 DRG 和阴道黏膜（VM）中的机制仍有待充分阐明。本研究调查了趋化因子 CXCL9、CXCL10 和 CXCL11 对 DRG 和 VM 中常驻 CD4 和 CD8 T 细胞的频率和功能的影响，以及其对复发性生殖器疱疹豚鼠模型中复发性生殖器疱疹的频率和严重程度的影响。HSV-2 潜伏感染的豚鼠通过肌肉内免疫 HSV-2 核糖核苷酸还原酶 2（RR2）蛋白（），随后通过阴道内给予表达趋化因子 CXCL9、CXCL10 或 CXCL11 的神经亲和性腺相关病毒 8 进行治疗，将 CD4 和 CD8 T 细胞募集到受感染的 DRG 和 VM（）。与 RR2 治疗性疫苗单独治疗相比，RR2/CXCL11 初免-拉动治疗性疫苗显著增加了 DRG 和 VM 组织中功能性组织驻留和效应记忆 CD4 和 CD8 T 细胞的频率。这与愈合的生殖器黏膜上皮中的病毒减少以及复发性生殖器疱疹的频率和严重程度降低有关。这些发现证实了局部 DRG 和 VM 常驻 CD4 和 CD8 T 细胞在减少感染部位阴道的病毒脱落和复发性生殖器疱疹严重程度方面的作用，并提出了初免-拉动疫苗策略来预防复发性生殖器疱疹。
重要性
本研究使用潜伏感染的豚鼠模型，调查了预防复发性生殖器疱疹的新型初免-拉动治疗性疫苗策略。在这项研究中，我们使用了一种策略，即使用重组表达单纯疱疹病毒 2 衣壳蛋白-核糖核苷酸还原酶 2（RR2；初免）免疫 HSV-2 感染的豚鼠，随后通过阴道内给予表达趋化因子 CXCL9、CXCL10 或 CXCL11 的神经亲和性腺相关病毒 8 将 T 细胞募集到受感染的背根神经节（DRG）和阴道黏膜（VM）（拉动）。我们表明，RR2/CXCL11 初免-拉动治疗性疫苗策略可显著减少阴道黏膜中的病毒脱落，并降低复发性生殖器疱疹的严重程度和频率。这种保护与潜伏感染的 DRG 组织和阴道黏膜愈合区域中浸润的功能性组织驻留（T 细胞）和效应（T 细胞）记忆 CD4 和 CD8 T 细胞的频率增加有关。这些发现揭示了组织驻留和效应记忆 CD4 和 CD8 T 细胞在 DRG 组织和 VM 中在预防复发性生殖器疱疹中的作用，并提出了初免-拉动治疗性疫苗策略来对抗生殖器疱疹。
